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Rui Kano

Publications and source records attributed to Rui Kano.

At least 19 recordsLinked to original sources
PubMed2006

An isolate of Prototheca wickerhamii from systemic canine protothecosis.

The isolate from a canine disseminated protothecosis was identified to be Prototheca wickerhamii by its morphological and biochemical characteristics. The isolate was grouped into a cluster identical to the type strain of P. wickerhamii, ATCC 16529(T) in phylogenetic trees based on the small subunit (SSU) and the 5' end of the large subunit (LSU) ribosomal DNA (rDNA); the cluster was close to that including the other Prototheca species. However, the strains of P. wickerhamii, SAG 263-11 and Pore 1283 belonged to a cluster different from the other isolates of Prototheca species and closely related to those of Auxenochorella species. Therefore, P. wickerhamii could be divided into two distinct genetic groups, one group close to the other Prototheca spp. including a standard strain of P. wickerhamii, and another group consisting of isolates previously reported to be close to the Auxenochorella species.

Animals↗
PubMed2006

Capsule-associated genes of serotypes of Cryptococcus neoformans, especially serotype AD.

Cryptococcus neoformans has been divided into five serotypes (A, B, C, D and AD) by the slide agglutination test against antigens of the polysaccharide capsule component. The isolates of serotype AD show positive reactions with both serotype A and D antigens. In this study, the nucleotide sequences of the capsule-associated genes CAP10, CAP59, CAP60 and CAP64 from five serotypes of C. neoformans were analyzed for their phylogenetic relationships, with special regard to serotype AD. The nucleotide sequence analyses showed that serotype AD had two different sequences in these four genes. Phylogenetic analysis revealed that these genes from serotype AD are included in the two clusters of serotypes A and D, but not in serotypes B or C. Southern blot analysis of genomic DNAs of serotypes A, D and AD digested with BamH I by hybridizing with a CAP64 probe indicated that both bands of fragments detected in serotypes A and D were also detected in serotype AD. These results confirm that serotype AD could be a mixture of serotypes A and D in the four CAP genes, consistent with its putative origin detected by the hybridization of these two serotypes. The present results indicated that serotypes of C. neoformans could be identified by the phylogenetic analyses of CAP genes.

Bacterial Capsules↗
PubMed2006

Apoptosis of canine mammary tumor cells induced by small interfering RNA (siRNA) against Bcl-xL gene.

The inhibition of Bcl-xL mRNA expression and the acceleration of apoptotic cell rates in canine mammary tumor cell line (CF33) by the small interfering RNA (siRNA) were analyzed. The level of Bcl-xL transcripts in CF33 was decreased when cultured with siRNA, suggesting that siRNA might inhibit the expression of Bcl-xL mRNA in the CF33. Apoptotic cell rates in CF33 cultured with siRNA in Oligofectamine medium, with double strand RNA in Oligofectamine medium, without siRNA in Oligofectamine medium and in DMEM alone were 60.9%, 30%, 28.7% and 11.6% at 48-hr incubation, respectively, when evaluated by TUNEL assay. From these results, it was suggested that canine Bcl-xL might be an anticancer target of canine tumors.

Animals↗
PubMed2006

A case of feline sporotrichosis.

We excised surgically a feline granulomatous lesion and performed histopathological, mycological and molecular examinations. As a result, it was diagnosed as sporotrichosis, which was the second recorded case of a cat so afflicted in Japan. After the operation, we recognized another nodule on the lymph node. Histopathological examination was therefore performed, but no fungi were detected. To prevent recurrence, the cat was administered a antimycotic drug, itraconazole. As a result, no recurrence was found. Excision of the lesion is the treatment of choice for feline sporotrichosis.

Animals↗
PubMed2006

Serotypes and mating types of clinical isolates from feline cryptococcosis in Japan.

Most isolates of Cryptococcus neoformans (teleomorph: Filobasidiella neoformans) from human patients and from environmental materials in Japan have been identified as serotype A mating type a by the seroagglutination test and mating experiments. A PCR method using the mating type alpha allele-specific primer of the STE12 gene and the serotype- and mating type-specific primers of the STE20 gene for identification of C. neoformans has been developed. Using the PCR method, conserved strains and clinical isolates from feline cryptococcosis were examined for serotype and the mating type. The results showed that all clinical isolates examined were identified as serotype A, MATalpha, indicating that feline cryptococcsis cases in Japan are caused by C. neoformans serotype A, MATalpha, as is the case in humans.

Animals↗
PubMed2005

Molecular cloning and tissue expression of canine Toll-like receptor 2 (TLR2).

Toll-like receptor 2 (TLR2) is involved in an important role on inflammatory reactions against various pathogenic microorganisms. Although some investigations suggested a relation between human and murine TLR2 gene polymorphisms and infections or inflammatory reactions, canine TLR2 gene has not been investigated. In the present study, therefore, the canine full-length of cDNA of Toll-like receptor 2 gene was sequenced and the expression of canine TLR2 mRNA was investigated in various tissues. Canine TLR2 clone was 2552bp encoding 785 amino acids. The similarities of canine TLR2 with human, mouse, rat, cattle, horse and pig TLR2 were 87.6, 72.9, 73.2, 83.1, 85.1 and 80.7% at the nucleotide sequence level, and 84.6, 68.7, 68.5, 78.8, 82.9 and 79.0% at amino acid sequence level, respectively. By a semiquantitative reverse transcription-polymerase chain reaction (RT-PCR) analysis, mRNA of canine TLR2 was detected in blood mononuclear cells (BMCs), lymph node, lung, liver, spleen, bladder, pancreas, small intestine, large intestine and skin of the dogs.

Amino Acid Sequence↗
PubMed2005

Canine CD20 gene.

The human CD20 antigen, a 35kDa cell surface nonglycosylated hydrophobic phoshpoprotein is expressed consistently on almost all human B-cells, and its monoclonal antibody is used for the therapy on human B-cell lymphoma. In the present study, canine CD20 gene was cloned and sequenced, and the expression of CD20 mRNA was investigated in canine peripheral blood mononuclear cells (PBMCs), and lymph nodes from healthy dogs, and canine lymphoma cells. Using canine cDNA as a template, full-length of canine CD20 gene was sequenced by 5'-RACE and 3'-RACE methods. The full-length of the cDNA sequence of canine CD20 was 1239bp encoding 297 amino acids. The amino acid sequences of canine CD20 showed 73 and 68% sequence similarities with those of human and mouse, respectively. Canine CD20 was predicted to contain domains of amino acid sequences consisting of two extracellular domains (EM), four transmembrane domains (TM), and three intracellular domains (IC) as in human CD20. Canine CD20 mRNA was detected in PBMCs and lymph node from healthy dogs, and B-cells of canine lymphoma, but not in T-cell lymphoma cells and non-T and non-B-cell lymphoma cells by RT-PCR analysis. From these results, canine CD20 might be targeted for monoclonal antibody therapy against B-cell lymphoma of dogs.

Amino Acid Sequence↗
PubMed2005

In vitro effect of recombinant human tumor necrosis factor-alpha on canine neutrophil apoptosis.

Neutrophils (polymorphonuclear leukocytes: PMNs) are essential for the host defense against various infections and are often injurious to the host, causing inflammatory diseases where tumor necrosis factor-alpha (TNF-alpha) is suggested to play an important role. Since an effect of TNF-alpha on canine PMN apoptosis has not been studied, canine PMNs were stimulated with recombinant human (rh)TNF-alpha in the present study to investigate the effect of TNF-alpha on canine PMN apoptosis. PMN apoptosis and function to produce ROS were assessed by flow cytometry. Delayed apoptosis was observed in the PMNs treated with rhTNF-alpha at 100 ng/ml, accompanied by retention of capability to produce ROS. However, PMN apoptosis was accelerated by rhTNF-alpha combined with cycloheximide. Therefore, it is indicated that TNF-alpha is able to activate anti- and pro-apoptotic pathways in PMNs and that the inhibition of PMN apoptosis by TNF-alpha requires protein synthesis in the PMNs.

Animals↗
PubMed2005

In vitro effect of recombinant human granulocyte colony-stimulating factor on canine neutrophil apoptosis.

Apoptosis is essential in eliminating neutrophils (polymorphonuclear leukocytes: PMNs) in animals. The suppression of PMN apoptosis is believed to be beneficial in eradicating pathogens and is implicated in the pathogenesis of human inflammatory diseases. In the present study, canine PMNs were stimulated with recombinant human granulocyte colony-stimulating factor (rhG-CSF) to investigate the in vitro effect on the apoptosis of canine PMNs. Apoptotic cell rates were assessed by flow cytometry in relation to the ability of PMNs to produce reactive oxygen species (ROS). Canine PMN apoptosis was markedly suppressed by rhG-CSF treatment, in association with the retention of the PMN ability to produce ROS. The addition of cycloheximide abolished this suppression by rhG-CSF. Moreover, canine PMNs, which were stimulated by rhG-CSF, expressed high levels of anti-apoptotic mcl-1 gene mRNA, as quantified by real-time polymerase chain reaction method. The results suggest that PMNs, stimulated by G-CSF, could work effectively over a longer period to eliminate pathogens, and that the prolongation of the PMN life-span might occasionally aggravate tissue injuries in dogs. In addition, the suppression of PMN apoptosis seems to be mediated by the induction of anti-apoptotic mcl-1 gene expression.

Animals↗
PubMed2005

Cloning of canine toll-like receptor 9 and its expression in dog tissues.

Toll-like receptor 9 (TLR9) is activated by bacterial DNA and induces production of inflammatory cytokines. In this study, canine TLR9 cDNA was cloned and sequenced. Further, the expression of TLR9 mRNA was investigated in canine tissues. The full-length cDNA for the canine TLR9 gene was 3419bp encoding 1032 amino acids. The similarity of canine TLR9 with those of cat, cattle, human, mouse and pig was 90, 84.3, 83.6, 76.3 and 85.2% at the nucleotide level, and 90.5, 82.7, 81.4, 74.8 and 85.7% at the amino acid level. By reverse transcription-polymerase chain reaction (RT-PCR) analysis, canine TLR9 mRNA was detected in venous blood mononuclear cells and lymph node and weakly in spleen, whereas it was not detected in kidney, liver, pancreas, lung, small intestine, large intestine, heart, skin, muscle, stomach and bladder.

Amino Acid Sequence↗
PubMed2005

Expression of capsule-associated genes of Cryptococcus neoformans.

Cryptococcus neoformans produces an extracellular polysaccharide capsule that is related to its virulence. The production of capsular components was reported to be accelerated when cultured on media with lower amount of glucose. In this study, relationship between capsule synthesis and expression of capsule-associated genes (CAP genes) was investigated by quantitative real-time PCR analysis. Normally encapsulated strains and a stable acapsular strain were cultured in 1% polypepton medium with 0.1% or 15% glucose. The results of assessment of the capsule size showed that the capsule of yeast cells cultured in the medium with low amount of glucose was thicker than that with high amount of glucose. The CAP gene expressions of normally encapsulated strains were higher in the medium with 0.1% glucose than in the medium with 15% glucose. Furthermore, CAP10, CAP59 and CAP60 genes were expressed very low in a stable acapsular strain, and CAP64 gene was not expressed. Results of assessment of capsule size and CAP gene expressions by quantitative real-time PCR analysis indicated that CAP gene expressions might be related to the production of capsule, and that glucose concentration in culture media might be related to the expression of CAP genes.

Cryptococcus neoformans↗
PubMed2005

Rapid identification of Candida tropicalis from canine cystitis.

The isolate from urine of a dog with cystitis was molecularly identified Candida tropicalis and its minimum inhibitory concentration (MIC) was determined by a microdilution method. The 25S ribosomal DNA sequence analysis indicated that the clinical isolate was essentially identical to that of C. tropicalis and distinct from other Candida species. The MIC(50) and the MIC(90) of fluconazole (FLZ) for the clinical isolate of C. tropicalis was 6.25 and 25 microg/ml, respectively, indicating that susceptibility of the clinical isolate of C. tropicalis to FLZ was less than for other strains of C. tropicalis as well as C. albicans. The molecular analysis as presented in this study assisted the diagnosis of candidiasis by identifying the yeasts in urine samples within 2 days. The patient dog, a 10-year-old male Shih Tzu dog (7.0 kg) referred for examination of cystitis was successfully treated with itraconazole.

Animals↗
PubMed2005

High expression of Bcl-xL in delayed apoptosis of canine neutrophils induced by lipopolysaccharide.

The expression of Bcl-2 family members, Bcl-xL, Bcl-2, Mcl-1 and Bax was investigated in delayed apoptosis of canine neutrophils induced by lipopolysaccharide (LPS). Apoptotic cell rates in neutrophils stimulated by LPS (100 ng/ml) were measured at 24 h incubation by TUNEL assay. The incidence of apoptotic neutrophils stimulated by LPS at 24 h incubation was 17.0+/-2% and that in non-stimulated neutrophils was 29.9+/-3%. By real-time quantitative PCR analysis, it was indicated that Bcl-xL and Bax levels in canine neutrophils were significantly affected by LPS stimulation. The levels of Bcl-xL, Bcl-2, Mcl-1 and Bax transcripts at 9 h incubation in neutrophils stimulated by LPS (100 ng/ml) were increased by about 80.4-, 1.9-, 1.4- and 5.3-folds, in comparison to those in non-stimulated neutrophils, respectively. These results indicated that Bcl-xL was proved have an important role in the inhibition of canine neutrophil apoptosis by LPS.

Animals↗
PubMed2005

Effect of antineoplastic drugs on the expression of Bcl-2 and Bcl-xL genes in the feline T-cell leukemia cell line.

The Bcl-2 gene is the first member of a rapidly expanding family of genes that regulate apoptosis. Bcl-2 has been shown to repress cell death triggered by a diverse array of stimuli including chemotherapy and gamma-irradiation. Chemotherapy of feline lymphoma is generally carried out with antineoplastic drugs, which are reported to induce apoptosis in tumor cells. However, the precise apoptotic signals, induced by chemotherapeutic drugs against feline tumors have not been fully characterized. Therefore, we have evaluated the expression of Bcl-2 and Bcl-xL in FT-1 upon in vitro treatment with these drugs. In the present study, full length of feline Bcl-xL gene was sequenced, and the expressions of Bcl-2 and Bcl-xL mRNAs in feline lymphoma cell line (FT-1) cultured with doxorubicin, prednisolone or vincristine were investigated. Feline Bcl-xL clone was 1163 base pairs in length and encoded 233 amino acids. The predicted amino acid sequence was 99.1%, 98.7%, 96.1%, 97.4%, 97.0% and 97.9% homologous to predicted Bcl-xL of dog, human, mouse, pig, rat and sheep, respectively. The levels of Bcl-2 transcripts at 24h incubation in FT-1 stimulated with doxorubicin (0.3mug/ml), prednisolone (0.2mug/ml) and vincristine (5ng/ml) were increased to about 41.0-, 62.0- and 11.1-fold to those in non-stimulated FT-1, respectively. On the other hand, the level of Bcl-xL transcripts at 24h incubation in FT-1 stimulated by doxorubicin and prednisolone were significantly increased about 4.2- and 5.8-folds to the controls and inducible level of Bcl-xL by vincristine was decreased about 0.35-folds.

Amino Acid Sequence↗
PubMed2005

Microsatellite polymorphism in Japanese mongrel dogs.

The genetic variability of 182 unrelated mongrel dogs living in various areas of Japan (from Hokkaido to Okinawa) was studied by collecting their blood. Ten microsatellite loci were chosen from different autosomal chromosomes. After combining a few rare adjoining alleles to allelic classes, it was confirmed that the Hardy-Weinberg equilibrium was attained in each locus. The polymorphic information contents (PICs) of the loci, Ren37A11, Ren48E01, AHTk253, ZuBeCa30, Ren277K09, Ren42N13, AHT130, PEZ03, PEZ12, and AHT121, were 0.58, 0.63, 0.67, 0.67, 0.68, 0.71, 0.79, 0.80, 0.80, and 0.80, and the power of discriminations (PDs) were 0.80, 0.85, 0.87, 0.88, 0.88, 0.89, 0.94, 0.94, 0.94, and 0.94, respectively. The combined mean exclusion chance (MEC) was 0.9995, indicating that these microsatellite loci are useful for kinship testing of Japanese dogs.

Animals↗